Cloning and Sequence Analysis of a Levansucrase Gene from Rahnella aquatilis ATCC15552

  • Kim, Hyun-Jin (Division of Biological Sciences, College of Natural Sciences, Pusan National University) ;
  • Yang, Ji-Young (Division of Food Science nd Biotechnology, Pukyung National University) ;
  • Lee, Hyeon-Gye (Department of Food and Nutrition, Hanyang University) ;
  • Cha, Jae-Ho (Division of Biological Sciences, College of Natural Sciences, Pusan National University)
  • Published : 2001.08.01

Abstract

An intracellular levansucrase gene, lscR from Rahnella aquatilis ATCC 15552, was cloned and its nucleotide sequence was determined. Nucleotide sequence analysis of this gene revealed a 1,238 bp open reading frame coding for a protein of 415 amino acids. The levansucrase was expressed by using a T7 promoter in Escherichia coli BL21 (DE3) and the enzyme activity was detected in the cytoplasmic fraction. The optimum pH and temperature of this enzyme for levan formation was pH 6 and $30^{\circ}C$, respectively. The deduced amino acid sequence of the lscR gene showed a high sequence similarity (59-89%) with Gram-negative levansucrses, while the level of similarity with Gram-positive enzymes was less than 42%. Multiple alignments of levansucrase sequences reported from Gram-negative and Gram-positive bacteria revealed seven conserved regions. A comparison of the catalytic properties and deduced amino acid sequence of lscR with those of other bacterial levansucrases strongly suggest that Gram-negative and Gram-positive levansucrases have an overall different structure, but they have a similar structure at the active site.

Keywords

References

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